一种用于定量检测细胞保护作用的快速灵敏的微量分析方法：用于分离抗参与脊髓灰质炎病毒附着的HeLa细胞蛋白的单克隆抗体

A rapid and sensitive micro scale assay for quantitative detection of cell protective effects: application for the isolation of a monoclonal antibody against HeLa cell proteins involved in poliovirus attachment.

作者信息

Barnert R H, Zeichhardt H, Habermehl K O

机构信息

Institut für Klinische und Experimentelle Virologie, Freie Universität Berlin, F.R.G.

出版信息

J Virol Methods. 1991 Nov;35(1):1-14. doi: 10.1016/0166-0934(91)90080-j.

DOI:10.1016/0166-0934(91)90080-j

PMID:1666110

Abstract

A combined assay consisting of a pre-cpe-protection assay and a double-antibody sandwich ELISA for detecting poliovirus was developed on a microtiter scale in order to quantify inhibition of virus replication caused by cell protective antibodies. The system was of high sensitivity and allowed the measurement of the protecting effect caused by a broad range of antibody concentrations before appearance of cytopathic effects. It was applied as a screening test for a large number of hybridomas secreting antibodies specific to the surface of HeLa cells and allowed the identification of four monoclonal antibodies (mAbs) with partial protection activity against poliovirus infection. One of the antibodies, mAb 122, detected SDS-PAGE-separated HeLa cell membrane proteins of 23-25 kDa and 50 kDa by immunoblot, indicating that these proteins are involved in poliovirus adsorption.

摘要

为了量化细胞保护抗体对病毒复制的抑制作用，在微量滴定板上开发了一种由前细胞病变效应保护试验和双抗体夹心酶联免疫吸附测定（ELISA）组成的联合检测方法，用于检测脊髓灰质炎病毒。该系统具有高灵敏度，能够在细胞病变效应出现之前测量广泛浓度抗体所产生的保护作用。它被用作一种筛选试验，用于检测大量分泌针对HeLa细胞表面特异性抗体的杂交瘤，并鉴定出四种对脊髓灰质炎病毒感染具有部分保护活性的单克隆抗体（mAb）。其中一种抗体mAb 122，通过免疫印迹法检测到SDS-PAGE分离的23 - 25 kDa和50 kDa的HeLa细胞膜蛋白，表明这些蛋白参与脊髓灰质炎病毒的吸附过程。